<?php echo"<div id='printVersion'><a href='../$protocolprint.php?csn=$csn&ssn=$ssn&dsn=$dsn&catno=$catno'>Print Version</a></div>";?>

<div class='MSDStitle'>Subculturing</div>
		<table width='100%'>
          <tr>
		    <td width='50' valign='top'></td>
			<td valign='top'>
             <?php
						   if($catno<>'T2009'){echo"
						      <ol>
								  <li>Remove and discard culture medium.</li>
								  <li>Add 2.0mL of Trypsin-EDTA solution (TM050) to flask and observe cells under an inverted microscope until cell layer is dispersed. Note: Cells that are  difficult to detach may be placed at 37°C to facilitate dispersal.</li>
								  <li>Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by  gently pipetting.</li>
								  <li>Centrifuge cells at 1500rpm for 3 minutes to pellet.</li>
								  <li>Aspirate out trypsin, leaving pellet undisturbed.</li>
								  <li>Resuspend pellet in fresh culture medium and plate in new culture vessel.</li>
								  <li>Incubate cultures at 37°C.</li>
                              </ol>";
						   }else{echo"These MEF cells were treated with 10μg/ml mitomycin C for 3 hours and can not proliferate anymore.<br/>
									Please thaw and plate the cells as follow:
									<ul style='margin-left:30px;'>
										<li>For 24-well plate: 5.0X10<sup>4</sup>-1.0X10<sup>5</sup> cells/ well</li>
										<li>For 6-well plate: 2.0X10<sup>5</sup>-4.0X10<sup>5</sup> cells/ well</li>
										<li>For 10 cm plate: 1.0X10<sup>6</sup>-2.5X10<sup>6</sup> cells</li>
										<li>For 15 cm plate: 5.0X10<sup>6</sup>-10.0X10<sup>6</sup> cells</li>
						   			</ul>";
						   }
			 ?>
            </td>
		  </tr>
		</table>
	  <div class='MSDStitle'>Preservation</div>
		<table>
          <tr>
		    <td width='50' valign='top'></td>
		    <td valign='top'>Freeze Medium: Complete growth medium with 20% FBS and 10% DMSO. Storage Temperature: Liquid nitrogen vapor phase.</td>
		  </tr>
		</table>